LOCUS       sequence-357210-        5380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HseIF4AI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5380)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5380)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pLNHA-C1-HseIF4AI_K
FEATURES             Location/Qualifiers
     source          1..5380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             665..730
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             755..781
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2168..2289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2205..2224)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2259..2278
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2296..2751)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2433..2454)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2645..2664
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2778..2882
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2884..3241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2884..3075
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2907..2927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3092..3227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3276..4070
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3276..4070
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3330..3349)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3940..3959
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4258..4277)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4302..4349
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4678..5266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5167..5186
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag ctcaatggaa
      721 agctgcaaac ggtggatctc ctccacatat ggcttaccca tacgatgttc cagattacgc
      781 tcctccatct cgagctcaag cttcgaattc tgcaatgtct gcgagccagg attcccgatc
      841 cagagacaat ggccccgatg ggatggagcc cgaaggcgtc atcgagagta actggaatga
      901 gattgttgac agctttgatg acatgaacct ctcggagtcc cttctccgtg gcatctacgc
      961 ctatggtttt gagaagccct ctgccatcca gcagcgagcc attctacctt gtatcaaggg
     1021 ttatgatgtg attgctcaag cccaatctgg gactgggaaa acggccacat ttgccatatc
     1081 aattctgcag cagattgaat tagatctaaa agccacccag gtcttggtcc tagcacccac
     1141 tcgagaattg gctcagcaga tacagaaggt ggtcatggca ctaggagact acatgggcgc
     1201 ctcctgtcac gcctgtatcg ggggcaccaa cgtgcgtgct gaggtgcaga aactgcagat
     1261 ggaagctccc cacatcatcg tgggtacccc tggccgtgta tttgatatgc ttaaccggag
     1321 atacctgtcc cccaaataca tcaagatgtt tgtactggat gaagctgacg aaatgttaag
     1381 ccgtggattc aaggaccaga tctatgacat attccaaaag ctcaacagca acacccaggt
     1441 agttttgctg tcagccacaa tgccttctga tgtgcttgag gtgaccaaga agttcatgag
     1501 ggaccccatt cggattcttg tcaagaagga agagttgacc ctggagggta tccgccagtt
     1561 ctacatcaac gtggaacgag aggagtggaa gctggacaca ctatgtgact tgtatgaaac
     1621 cctgaccatc acccaggcag tcatcttcat caacacccgg aggaaggtgg actggctcac
     1681 cgagaagatg catgctcgag atttcactgt atccgccatg catggagata tggaccaaaa
     1741 ggaacgagac gtgattatga gggagtttcg ttctggctct agcagagttt tgattaccac
     1801 tgacctgctg gccagaggca ttgatgtgca gcaggtttct ttagtcatca actatgacct
     1861 tcccaccaac agggaaaact atatccacag aatcggtcga ggtggacggt ttggccgtaa
     1921 aggtgtggct attaacatgg tgacagaaga agacaagagg actcttcgag acattgagac
     1981 cttctacaac acctccattg aggaaatgcc cctcaatgtt gctgacctca tctgaccggg
     2041 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     2101 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2161 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2221 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2281 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2341 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2401 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2461 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2521 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2581 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2641 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2701 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     2761 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2821 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2881 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2941 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3001 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3061 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3121 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3181 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3241 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3301 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3361 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3421 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     3481 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3541 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3601 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3661 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3721 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3781 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     3841 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3901 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3961 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4021 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4081 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4141 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4201 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     4261 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4321 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4381 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4441 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4501 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4561 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4621 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4681 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4741 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4801 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4861 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4921 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4981 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5041 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5101 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5161 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5221 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5281 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5341 atcccctgat tctgtggata accgtattac cgccatgcat
//
